2.1 Fluorescent Microscope

Question 1

Visible light

Answer 1

400 nm - 700 nm

Question 2

Fluorescence

Answer 2

Specific molecules located in cells by fluorescence microscopy

antibodies used to detect specific molecules.

Question 3

Fluorescent Tags

Answer 3

fluorescent proteins used to tag individual proteins - living cells + organisms

protein dynamics followed in living cells

Question 4

Immunocytochemistry + Immunofluorescence

Answer 4

primary antibodies:
rabbit antibody directed against antigen A

secondary antibodies:
marker-couples antibodies directed against rabbit antibodies

1. first barrier filter:
   lets through only blue light with wavelength between 450 nm + 490 nm
2. beam splitting mirror:
   reflects light below 510 nm - transmits light above 510 nm
3. second barrier filter:
   cuts out unwanted fluorescent signals - passes specific green fluorescein emission between 520 nm + 560 nm

Question 5

Principles of confocal microscopy

Answer 5

A) fluorescent specimen is illuminated with focused light point from pinhole

B) emitted fluorescent light from in-focus point focused at pinhole + reaches detector

C) emitted light from out-of-focus point is out of focus st pinhole + largely excluded from detector

Question 6

Photoactivated fluorescent dyes

Answer 6

0 second - photoactivation
1 second - fluorescence in selected region
4 minutes
13 minutes